Dynamics of proteins in Golgi membranes: comparisons between mammalian and plant cells highlighted by photobleaching techniques

In less than a decade the green fluorescent protein (GFP) has become one of the most popular tools for cell biologists for the study of dynamic processes in vivo. GFP has revolutionised the scientific approach for the study of vital organelles, such as the Golgi apparatus. As Golgi proteins can be tagged with GFP, in most cases without altering their targeting and function, it is a great substitute to conventional dyes used in the past to highlight this compartment. In this review, we cover the application of GFP and its spectral derivatives in the study of Golgi dynamics in mammalian and plant cells. In particular, we focus on the technique of selective photobleaching known as fluorescence recovery after photobleaching, which has successfully shed light on essential differences in the biology of the Golgi apparatus in mammalian and plant cells.     

芽孢杆菌植酸酶基因在毕赤酵母中的胞内表达

根据已知的枯草芽孢杆菌WHNB02植酸酶phyC基因全序列设计一对引物,采用PCR法从含有该基因的pUC18-phyC质粒上获得了长约1.1kb不含有信号肽序列的植酸酶phyC基因表达片段.经T载体克隆及序列测定后,构建毕赤巴斯德表达载体pPIC3.5K-phyC,并电转化毕赤巴斯德酵母宿主菌GS115.经MD和MM平板筛选、酶活性测定,获得了阳性转化子,并进行了诱导表达.SDS-PAGE分析表明:表达产物分子量为42.01KD,表达量占细胞可溶性总蛋白的24%,并具有植酸酶的生物学活性.酶学性质分析结果显示:胞内表达的植酸酶酶促反应最适pH值为7.5;最适反应温度为70℃;经90℃处理10min,残留酶活性42% 均优于出发菌株天然植酸酶的相应性质.     

Icariin suppresses bone resorption activity of rabbit osteoclasts in vitro

The effect of icariin on the bone resorption activity of rabbit osteoclasts is assessed in vitro. Osteoclasts were isolated from Japanese white rabbits and cultured on plates with a sterilized bone slice in each well. After treatment with icariin at various concentrations, the bone resorption activity of osteoclasts was evaluated by examining pit areas, superoxide anion (·O2-) generation, size and number of actin rings and intracellular calcium concentration [Ca2 ]i. As revealed by these data, icariin elicited continuous decline of [Ca2 ]i, making actin ring constricted and ·O2- generation decreased. These events resulted in smaller and fewer pits which indicate suppressed bone resorption activity of rabbit osteoclasts by icariin.     

Depolarization Induced by Substance P on Rat Stellate Ganglion Neurons

应用细胞内生物电记录技术，观察神经肽Ｐ物质（ＳＰ）对大鼠星状神经节细胞的影响。ＳＰ在１μｍｏｌ～１０μｍｏｌ或更高的浓度范围内，供试３５个细胞，有２８个细胞发生膜除极反应。用低钙（０．２５ｍｍ）或用含河豚毒素（ＴＴＸ，１μｍｏｌ）克氏液灌流神经节，不影响ＳＰ引起的除极反应的幅度和时程。ＳＰ引起除极反应的同时常伴有膜电阻增大。当膜电位增大时，除极化反应幅度变小，反转电位为－８０ｍＶ至－１００ｍＶ。研究表明，ＳＰ对部分星状神经节细胞具有兴奋作用，使通过这些细胞的信息传递增强；ＳＰ对细胞膜的除极作用是由于其引起细胞膜钾导降低所致。     

Cholera toxin structure, gene regulation and pathophysiological and immunological aspects

Many notions regarding the function, structure and regulation of cholera toxin expression have remained essentially unaltered in the last 15 years. At the same time, recent findings have generated additional perspectives. For example, the cholera toxin genes are now known to be carried by a non-lytic bacteriophage, a previously unsuspected condition. Understanding of how the expression of cholera toxin genes is controlled by the bacterium at the molecular level has advanced significantly and relationships with cell-density-associated (quorum-sensing) responses have recently been discovered. Regarding the cell intoxication process, the mode of entry and intracellular transport of cholera toxin are becoming clearer. In the immunological field, the strong oral immunogenicity of the non-toxic B subunit of cholera toxin (CTB) has been exploited in the development of a now widely licensed oral cholera vaccine. Additionally, CTB has been shown to induce tolerance against co-administered (linked) foreign antigens in some autoimmune and allergic diseases. Received 25 October 2007; accepted 12 December 2007     

Huntington’s disease: from huntingtin function and dysfunction to therapeutic strategies

Huntington’s disease (HD) is a neurodegenerative disorder that usually starts in middle age and is characterized by involuntary movements (chorea), personality changes and dementia, leading to death within 10–20 years. The defective gene in HD contains a trinucleotide CAG repeat expansion within its coding region that expresses a polyglutamine repeat in the protein huntingtin. Together with the characteristic formation of aggregates in HD, aberrant protein interactions and several post-translational modifications affect huntingtin during disease progression and lead to the dysfunction and death of selective neurons in the brains of patients. The exact molecular mechanisms by which mutant huntingtin induces cell death are not completely understood but may involve the gain of new toxic functions and the loss of the beneficial properties of huntingtin. This review focuses on the cellular functions in which huntingtin is involved and how a better understanding of pathogenic pathways can lead to new therapeutic approaches. Received 24 May 2006; received after revision 5 July 2006; accepted 23 August 2006     

Effect of Ab_(1-40) on membrane permeability and intracellular free Ca~(2+) of nerve cells

Alzheimers disease (AD) is a kind of progressive neurodegenerative disease, characterized by intracellularfibrillar tangles and extracellular senile plaques, the main component of the plaques are b-amyloid (Ab), composed of 3943 amino acids[1]. Glenner and Wong first isolated Ab peptide and determined the sequence in 1984, after-wards, a series of researches were done on its production, structure determination, aggregation, hydrophobicity, re-ceptor and cell toxicity. Ab is the abnormal fiss…     

基于视频特征聚合的细胞形变动态建模

提出了一种基于图像处理架构对活细胞视频中细胞动态进行建模的方法.在这个架构下,视频中两帧间的细胞动态被表示为细胞轮廓形变和细胞内部运动的动态特征.其中,前者细胞轮廓形变利用形状上下文进行度量,而后者细胞内部运动则通过尺度不变特征变换（SIFT）流进行建模.为了更好的刻画细胞质流动,基于SIFT流的细胞运动场进一步构建细胞外观变化场.在获得上述帧级细胞动态特征之后,引入时间序列建模的方法来生成视频级的细胞动态特征.具体地,基于紧凑编码的时序特征聚合方法可以捕获整个视频中的细胞动态演变过程.活细胞视频数据库被建立以用于验证提出方法的有效性,实验结果表明提出方法对于细胞形变动态度量和分类的性能优于其它主流方法.     

Effects of pCai and pHi on cell-to-cell coupling

Summary Internal longitudinal resistance (r_i), a determinant of cardiac conduction, is affected by changes in intracellular calcium and protons. However, the role and mechanism by which H⁺ and Ca²⁺ may modulate r_i is uncertain. Cable analysis was performed in cardiac Purkinje fibers to measure r_i during various interventions. In some experiments, intracellular pH (pH_i) was recorded simultaneously to study the pH_i-r_i relation. Both intracellular Ca²⁺ and H⁺ independently modified r_i. However, internal resistance of cardiac fibers was insensitive to pH_i changes compared to other tissues. A latent period preceded the pH_i-related changes in r_i and the amount of change depended upon methodology. The results suggest that direct action of protons on r_i may be subordinate to other regulatory processes. Ionic regulation of internal longitudinal resistance may occur by more than one mechanism: i) direct cationic binding to sites on junctional membrane proteins; and ii) H⁺- or Ca²⁺-dependent phosphorylation of junctional proteins.     

骨桥蛋白在免疫中的作用

骨桥蛋白（OPN）是一种多功能的磷酸化糖蛋白,其涉及多种病理生理学过程.近来研究发现骨桥蛋白通过不同的翻译起始位点产生两种形式的蛋白,即分泌型骨桥蛋白（ sOPN,secreted OPN）和胞内型骨桥蛋白（iOPN,intracellular OPN）.本文主要介绍了sOPN及iOPN在免疫应答中的作用.